Characterization of Defective Interfering RNA Components That Increase Symptom Severity of Broad Bean Mottle Virus Infections

Virology ◽  
1993 ◽  
Vol 194 (2) ◽  
pp. 576-584 ◽  
Author(s):  
Javier Romero ◽  
Qi Huang ◽  
Judit Pogany ◽  
Jozef J. Bujarski
1960 ◽  
Vol 15 (7) ◽  
pp. 444-447 ◽  
Author(s):  
C. Wetter ◽  
H. L. Paul ◽  
J. Brandes ◽  
L. Quantz

Broad bean true mosaic virus (EAMV) has been studied in comparison with broad bean mottle virus (BBMV), after both viruses had been purified by the same procedure. Spectrophotometric and/or chemical determinations revealed a nucleic acid content of 32% for EAMV and 23% for BBMV. Serologically, the two viruses are unrelated since in reciprocal testings they react with their homologous antisera only. The diameter of of the particles has been determined to 25 mμ for EAMV and 20 mμ for BBMV. Further differences include thermal inactivation point, seed transmission, host range, and symptomatology.There are no indications for a relationship among both viruses as suggested in the list of Common names (Review of Applied Mycology, Vol. 35, Suppl.).


2000 ◽  
Vol 74 (7) ◽  
pp. 3156-3165 ◽  
Author(s):  
Richard Molenkamp ◽  
Babette C. D. Rozier ◽  
Sophie Greve ◽  
Willy J. M. Spaan ◽  
Eric J. Snijder

ABSTRACT Equine arteritis virus (EAV), the type member of the family Arteriviridae, is a single-stranded RNA virus with a positive-stranded genome of approximately 13 kb. EAV uses a discontinuous transcription mechanism to produce a nested set of six subgenomic mRNAs from which its structural genes are expressed. We have generated the first documented arterivirus defective interfering (DI) RNAs by serial undiluted passaging of a wild-type EAV stock in BHK-21 cells. A cDNA copy of the smallest DI RNA (5.6 kb) was cloned. Upon transfection into EAV-infected BHK-21 cells, transcripts derived from this clone (pEDI) were replicated and packaged. Sequencing of pEDI revealed that the DI RNA was composed of three segments of the EAV genome (nucleotides 1 to 1057, 1388 to 1684, and 8530 to 12704) which were fused in frame with respect to the replicase reading frame. Remarkably, this DI RNA has retained all of the sequences encoding the structural proteins. By insertion of the chloramphenicol acetyltransferase reporter gene in the DI RNA genome, we were able to delimitate the sequences required for replication/DI-based transcription and packaging of EAV DI RNAs and to reduce the maximal size of a replication-competent EAV DI RNA to approximately 3 kb.


1961 ◽  
Vol 16 (12) ◽  
pp. 786-791 ◽  
Author(s):  
H. L. Paul

Broad bean mottle virus (BBMV) contains 15.2 percent N and 1.83 percent P; the N/P ratio is 8.4 ± 0.3. The protein of BBMV contains 15.6 percent N and no P; the NA has a N/P ratio of about 1.85. From these data a NA content of about 22 percent can be evaluated. This value agrees with that of 23 percent obtained by orcinol reactions.The sedimentation constant at infinite dilution is 83 ± 2 S, the diffusion constant 1.44 ± 0.04 · 10-7 cm2/sec, and the partial specific volume 0.75 cm3/gram. From these data a molecular weight of 5.6 ± 0.5 millions for the unhydrated particles can be calculated. The diameter of these particles is about 24 mµ, which agrees fairly well with determinations made with the electron microscope 1. Estimations of molecular weight and diameter of the hydrated particles reveal about 10 millions and about 30 mu respectively.Purified BBMV was stable in neutral buffer solutions at 3 °C for some weeks and remained infective. Exhaustive dialysis of such virus suspensions against dist. water caused a considerable increase in turbidity, and, later on, precipitation of the virus. It was not possible to resuspend this virus completely by adding neutral buffer or salt solutions. Density gradient centrifugations showed these resuspensions to be inhomogeneous, whereas non-dialyzed virus suspensions in buffer gave only one narrow zone in the gradient tubes, indicating the homogeneity of the particles.In buffers of different pH , virus suspensions showed a maximum turbidity at about pH 4.5. For precipitations of BBMV with ammonium sulfate, higher salt concentrations were necessary than for most other viruses.The UV light absorption (corrected for light scattering) of BBMV as well as of its protein and its NA was measured. It could be shown that the absorption coefficient of BBMV protein is much lower than that of most other viruses, presumably because of the low content of aromatic amino acids as determined by chemical analysis recently 2.The estimation of the NA content of BBMV from UV absorption data is discussed.


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